6 replies to this topic

[microbes]

Hello
My Department (India) want to purchase a new RT PCR, options are

Illumina   Eco real Time 4 emission filters
ABI Step One    3 filters
Roche Light Cycler 480    6 filters

48 wells will serve the purpose,
I do not know much about these systems.
are extra filters (4/6) of much significance?
Is it that Roche system require chemicals of Roche also, than other company?

Thanks for suggestions..

Edited by microbes, 20 January 2012 - 10:43 AM.

[Adrian K]

If you want a multipurpose machine where you can have most of the applications, Roche 480 is a good one, else Rotogene 6000.
If you want entry level, Try Roche Nano, and Illumina Eco.
Roche system compatible with other chemicals, even for homebrew mastermix.

[Trof]

Step One or LightCycler are fine machines. LC480 is more multipurpose (HRM, probe melting, etc.) Step One is cheaper I think and smaller, and don't need separate computer. But I would really consider the 48 wells. It may look enough, but can be limiting. You run a dillution curve with your sampes and all triplicates and you can only have 11 samples. ABI offers Step One Plus that has 96 wells. You can often run more than one gene on one plate, if they're designed the same way.

LC480 has the disadvantage in need to use plates, not individual tubes or strips. If you really have only small sample sizes this could be wasting too much, as you can use the plate (unused positions) only twice, more than that gives ugly curves. Step One systems can use anything.

LC480 has more filters which means wider range of fluorophores, that you don't usually need. For quantification you need FAM/SYBR and VIC, that's two filters. All machines can use any chemistry, but the representatives will tell you theirs work the best (this is true for LC480 and Roche mixes as we we able to try).

[microbes]

thanks for the suggestions...

we have planned to buy ABI step one. it can be used as a PCR also but not other machines..and i think 3 filters with common dyes will be enough and 48 wells can be best to start with, and can be upgraded to 96 later when required. this machine has maximum installations in my region as well as country.
frankly speaking I still have to learn about what experiments to do actually with this machine, and hope the actual buyer will be in good chemistry with my group..

[Adrian K]

Having one StepOne and later upgraded to stepone plus in the department, I will not recommend you to get this machine.

1) As far as I understand, You need to purchase ABI plates, strips, seal etc in order to use StepOne system. You unable to use other brand's (Axygen etc..) for StepOne. They are using 0.1ml strips which other manufacturer's do not fit, last time I try (I have to admit that is very long time ago). So, using StepOne as normal PCR machine will increase your research cost.

2) Agree that StepOne you can program your run on the qPCR machine itself without the need of additional computer, and thats the only thing that you can do. After your qPCR run, and if you do not save your run data immediately, you will lost all your running data if someone else start their run and did not save yours, as there is no internal memory for you to do so. (I would say this is a very stupid design by ABI, your need to end up plugging your thumbdrive on the machine when you do your run). You can install the StepOne client on your own lab PC for your qPCR configuration and data analysis. FYI, HRM analysis requires you to purchase another software (dongle) in order to run the HRM, else it will be handicapped.

Think twice before you get this StepOne  machine, I totally not recommend it. You can try stratagene MX3000, I "heard" (never use before) is a very good one.

[Trof]

I bet there are other companies offering plastics for StepOne, I found Eurogentec for example. But of course manufacturer will always tell you you shouldn't use it, that's on one's risk always, they may be right or not.

Adrian, did you try to connect the machine to the network? They told us it could be used this way, are the data accessible this way or downloaded automatically or not?

Just noted, for LC480 you need to buy HRM analysis module also separately.

[Adrian K]

Wow... I should inform my colleagues look for the local distributor for Eurogentec now... thanks

Trof, I want to try that feature but however, we are not authorized to connect it to the network, as according to our department , they fear that someone might abused it, computer virus infections etc. But I do believe they have hidden agenda.

LC480 need separate HRM module as well? Geez, thats bad. Roche should bundle it once you guys buy the LC480 system.